Different RNA and protein expression of surface markers in rabbit amniotic fluid-derived mesenchymal stem cells

Authors

  • Michal Kovac,

    Corresponding author
    1. Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic
    2. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
    • Correspondence concerning this article should be addressed to M. Kovac at 3mkovac3@gmail.com.

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  • Jaromir Vasicek,

    1. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
    2. Research Centre AgroBioTech, Slovak University of Agriculture, Nitra, Slovak Republic
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  • Barbora Kulikova,

    1. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
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  • Miroslav Bauer,

    1. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
    2. Faculty of Natural Sciences, Constantine the Philosopher University, Nitra, Slovak republic
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  • Jozef Curlej,

    1. Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic
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  • Andrej Balazi,

    1. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
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  • Peter Chrenek

    1. Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic
    2. Research Inst. for Animal Production, National Agricultural and Food Centre, Lužianky, Slovak Republic
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Abstract

Over the years, there has been much confusion in defining molecular markers of mesenchymal stem cells (MSCs) for other than human species due to a lack of species-specific antibodies. Therefore, the aim of our study was to define rabbit amniotic fluid-derived mesenchymal stem cells (rAF-MSCs) and to reflect upon the current identification of AF-MSCs by providing a summary of detected surface markers in different species. The expression of rAF-MSC surface markers was analyzed at the protein and mRNA level. Flow cytometry analyses showed that rAF-MSCs were positive for CD29 and CD44, low positive for CD90, but negative for CD73, CD105, and CD166. Interestingly, RT-PCR (reverse transcription-polymerase chain reaction) exposed a discprepancy between transcribed mRNA and protein expression, as the cells expressed mRNA of all MSC markers: CD29, CD44, CD73, CD90, CD105, and CD166. Our results also confirmed the mesenchymal nature of isolated cells by morphology, ultrastructure, and intracellular marker expression profile. In addition, the expression of few pluripotency markers was also detected. We also found that passaging did not affect apoptosis and viability. Similarly, changes in karyotype were not observed during passaging. In conclusion, the provided characteristics may be used as a comprehensive set of criteria to define and characterize rAF-MSCs, required for the identification of these cells in preclinical investigations. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1601–1613, 2017

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